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Bovine oocyte plasma membrane binding sites for sperm plasma membrane during in vitro oocyte maturation and fertilisation

Published online by Cambridge University Press:  26 September 2008

Emily Wheeler ,
Trish Berger  and
Esmail Behboodi
Emily Wheeler
Affiliation:
Department of Animal Science, University of California, Davis, California, USA
Trish Berger*
Affiliation:
Department of Animal Science, University of California, Davis, California, USA
Esmail Behboodi
Affiliation:
Department of Animal Science, University of California, Davis, California, USA
*
Trish Berger, Department of Animal Science, University of California Davis, Davis, CA 95616-8521, USA. Telephone: +1 (916) 752-1267. Fax: +1 (916)752-0175.
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Summary

The experimental objective was to determine whether the capability of bovine oocyte plasma membrane to bind sperm changes during in vitro oocyte maturation and fertilisation. Binding was quantified by the intensity of tetramethylrhodamine isothiocyanate (TRITC) fluorescence at the periphery of oocytes following incubation with biotinylated sperm plasma membrane proteins and subsequent incubation with TRITC-avidin. Bovine oocytes were matured in vitro. Sample groups were removed after 0,6 and 22 h, or inseminated and further cultured for 24 or 48 h. Oocytes were denuded of cumulus cells and zona pellucida and co-incubated with 56 μg biotinylated bovine sperm plasma membrane protein for 45 min in 150 μl drops of saline-BSA. Controls were incubated for the same time period in the absence of sperm plasma membrane proteins. All oocytes were rinsed, incubated with TRITC-avidin and subsequently fixed and transferred to mounting medium. Oocytes were scanned with a confocal microscope and analysed using ImageQuant software. The binding of sperm plasma membrane was quantified by integrated fluorescent intensity in standardised ellipses spaced around the plasma membrane of the oocyte. Values are expressed as mean intensity units per 320 pixel ellipse. Binding of sperm plasma membrane continued to increase throughout in vitro oocyte maturation and fertilisation (9051, 24318 and 49953 for 0 and 22 h in vitro matured oocytes and fertilised oocytes, respectively; p = 0.0001). A dramatic decrease in sperm plasma membrane binding to the oocyte plasma membrane was observed in 2-cell embryos (mean intensity = 24477, p = 0.0001). The observed binding was primarily due to the binding of sperm plasma membrane proteins, as control oocytes incubated with TRITC- avidin only were barely visible (integrated fluorescence intensity values ranged from 8 to 3757).

Keywords

Egg plasma membraneEmbryoOolemmaSperm–egg interaction
Type
Article
Information
Zygote , Volume 4 , Issue 1 , February 1996 , pp. 67 - 72
Copyright
Copyright © Cambridge University Press 1996

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References

Almeida, E.A.C., Huovila, A.P.J., Sutherland, A.E., Stephens, L.E., Calarco, P.G., Shaw, L.M., Mercurio, A.M., Sonnen-berg, A., Primakoff, P., Myles, D.G. & White, J.M. (1995). Mouse egg integrin α6β1 functions as a sperm receptor. Cell 81, 1095–104.CrossRefGoogle ScholarPubMed
Ash, K., Berger, T., Munn, R.J. & Horner, C.M. (1994). Isolation and partial purification of plasma membrane from porcine oocytes. Mol. Reprod. Dev. 38, 334–7.CrossRefGoogle ScholarPubMed
Ash, K., Berger, T., Horner, C.M. & Calvert, C.C. (1995). Identification of porcine sperm plasma membrane proteins that may play a role in sperm-egg fusion. Zygote 3, 163–70.CrossRefGoogle ScholarPubMed
Behboodi, E., Anderson, G.B. & BonDurant, R.H. (1992). Development of in vitro fertilized oocytes from pregnant and nonpregnant cows in oviductal epithelial and cumulus cell co-culture systems. Theriogeniology 38, 1077–84.CrossRefGoogle ScholarPubMed
Behboodi, E., Anderson, G.B., Horvat, S., Medrano, J.F., Murray, J.D. & Rowe, J.D. (1993). Microinjection of bovine embryos with a foreign gene and its detection at the blastocyst stage. J. Dairy Sci. 76, 3392–9.CrossRefGoogle ScholarPubMed
Blobel, C.P., Wolfsberg, T.G., Turck, C.W., Myles, D.G., Primakoff, P. & White, J.M. (1992). A potential fusion peptide and an integrin ligand domain in a protein active in sperm-egg fusion. Nature 356, 248–52.CrossRefGoogle Scholar
Cran, D.G. & Moor, R.M. (1989). Programming the oocyte for fertilization. In Fertilization in Mammals, ed. Bavister, B.D., Cummins, J. & Roldan, E.R.S., pp. 35–47. Norwell, Mass.: Serono Symposium.Google Scholar
Gillis, G., Peterson, R., Russell, L., Hook, L. & Freund, M. (1978). Isolation and characterization of membrane vesicles from human and boar spermatozoa: methods using nitrogen cavitation and ionophore induced vesiculation. Prep. Biochem. 8, 363–78.Google ScholarPubMed
Kellom, T., Vick, A. & Boldt, J. (1992). Recovery of penetration ability in protease-treated zona-free mouse eggs occurs coincident with recovery of a cell surface 94 kD protein. Mol. Reprod. Dev. 33, 46–52.CrossRefGoogle ScholarPubMed
Moor, R.M., Mattioli, M., Ding, J. & Nagai, T. (1990). Maturation of pig oocytes in vivo and in vitro. J. Reprod. Fertil. Suppl. 40, 197–210.Google ScholarPubMed
Myles, D.G., Kimmel, L.H., Blobel, C.P., White, J.M. & Primakoff, P. (1994). Identification of a binding site in the disintegrin domain of fertilin required for sperm-egg fusion. Proc. Natl. Acad. Sci. USA 91, 4195–8.CrossRefGoogle ScholarPubMed
Okabe, M., Matzno, S., Nagira, M., Mimura, T., Kawai, Y. & Mayumi, T. (1990). A human sperm antigen possibly involved in binding and/or fusion with zona-free hamster eggs. Biol. Reprod. 30, 731–6.Google Scholar
O'Rand, M.G. & Irons, G. (1984). Monoclonal antibodies to rabbit sperm autoantigens. II. Inhibition of human sperm penetration of zona-free hamster eggs. Biol. Reprod. 30, 731–6.CrossRefGoogle ScholarPubMed
Parrish, J.J., Susko-Parrish, J.L., Weiner, M.A. & First, N.L. (1988). Capacitation of bovine spermatozoa by heparin. Biol. Reprod. 38, 715–20.CrossRefGoogle ScholarPubMed
Peterson, R., Russell, L., Bundman, D. & Freund, M. (1980). Evaluation of the purity of boar sperm plasma membranes prepared by nitrogen cavitation. Biol. Reprod. 23, 637–45.CrossRefGoogle ScholarPubMed
Primakoff, P., Hyatt, H. & Tredick-Kline, J. (1987). Identification and purification of a sperm surface protein with a potential role in sperm-egg membrane fusion. J. Cell Biol. 104, 141–9.CrossRefGoogle ScholarPubMed
Rochwerger, L., Cohen, D.J. & Cuasnicu, P.S. (1992). Mammalian sperm-egg fusion: the rat egg has complementary sites for a sperm protein that mediates gamete fusion. Dev. Biol. 153, 83–90.CrossRefGoogle ScholarPubMed
Saling, P.M., Raines, L.M. & O'Rand, M.G. (1983). Monoclonal antibody against mouse sperm blocks a specific event in the fertilization process. J. Exp. Zool. 227, 481–6.CrossRefGoogle ScholarPubMed
Taylor, C.T., Biljan, M.M., Kingsland, C.R. & Johnson, P.M. (1994). Inhibition of human spermatozoan-oocyte interaction in vitro by monoclonal antibodies to CD46 (membrane cofactor protein). Hum. Reprod. 9, 907–11.CrossRefGoogle ScholarPubMed
Wasserman, P.M. (1994). Gamete interactions during mammalian fertilization. Theriogeniology 41, 31–44.CrossRefGoogle Scholar
Yanagimachi, R. (1994). Mammalian fertilization. In The Physiology of Reproduction, ed. Knobil, E. & Neill, J.D., 2nd edn, pp. 189–317. New York: Raven Press.Google Scholar
Zuccotti, M., Yanagimachi, R. & Yanagimachi, H. (1991). The ability of hamster oolemma to fuse with spermatozoa: its acquisition during oogenesis and loss after fertilization. Development 112, 143–52.CrossRefGoogle ScholarPubMed